网络出版日期: 2021-12-03
基金资助
2018年度河南省医学科技攻关计划项目(2018020950)
Effects of knockdown of immunoregulatory protein B7-H3 gene on malignant proliferation, invasion and stem cell-like characteristics of bladder cancer cells
Online published: 2021-12-03
Supported by
2018 Henan Province Medical Science and Technology Tackling Project(2018020950)
目的·探讨干扰免疫调节蛋白B7-H3基因对膀胱癌J82细胞的恶性增殖和干细胞样特性的影响。方法·shRNA干扰效率的检测采用反转录聚合酶链式反应（reverse transcription-polymerase chain reaction，RT-PCR）。J82细胞增殖采用克隆形成实验检测。细胞凋亡采用流式细胞术检测。细胞侵袭采用Transwell检测。肿瘤克隆能力采用肿瘤成球实验观察。细胞凋亡、侵袭和干细胞样相关蛋白表达采用蛋白质印迹法（Western blotting）检测。结果·B7-H3在膀胱癌组织及膀胱癌细胞系中高表达（P=0.000）。确定shRNA2为设计的3组shRNAs中干扰B7-H3基因效率最高（P=0.000）。B7-H3干扰抑制了J82细胞的增殖、侵袭、肿瘤克隆能力及干细胞样特性（P=0.000）。并且，B7-H3干扰促进了J82细胞的凋亡（P=0.000）。结论·免疫调节蛋白B7-H3基因干扰可对膀胱癌J82细胞的恶性增殖、侵袭及干细胞样特性产生抑制作用。
王晓寒 , 叶云林 , 肖康华 , 赵志鑫 , 吴静雅 . 敲减免疫调节蛋白B7-H3基因对膀胱癌细胞恶性增殖、侵袭和干细胞样特性的影响[J]. 上海交通大学学报（医学版）, 2021 , 41(11) : 1454 -1460 . DOI: 10.3969/j.issn.1674-8115.2021.11.008
·To investigate the effect of gene interfering with immune regulatory protein B7-H3 on malignant proliferation and stemness characteristics of bladder cancer J82 cells.
·Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect shRNA interference efficiency. The proliferation of J82 cells was detected by clone formation assay. Cell apoptosis was detected by flow cytometry. Cell invasion was detected using Transwell asssay. The tumor-cloning ability was observed using tumor sphere formation experiment. Apoptosis, invasion, and expression and stemness-related proteins were detected by Western blotting.
·B7-H3 is highly expressed in bladder cancer tissues and bladder cancer cell lines (P=0.000). shRNA2 was identified as the most efficient interference with B7-H3 gene in three shRNAs (P=0.000). The interference of B7-H3 inhibited the proliferation, invasion, tumor cloning ability and stem cell-like characteristics of J82 cells (P=0.000). Furthermore, B7-H3 interference promoted the apoptosis of J82 cells (P=0.000).
·The gene interference of immunoregulatory protein B7-H3 can inhibit the malignant proliferation, invasion and stemness characteristics of bladder cancer J82 cells, providing a new possibility and effective evidence for the treatment of bladder cancer.
Key words： B7-H3; Bladder cancer; J82 cells; Stemness characteristics
| 1 | Sanli O, Dobruch J, Knowles MA, et al. Bladder cancer[J]. Nat Rev Dis Primers, 2017, 3: 17022. |
| 2 | 白志鹏, 唐以众. TURBT联合表柔比星与羟喜树碱膀胱灌注对浅表性膀胱癌患者术后复发率及生活质量的影响[J]. 解放军医药杂志, 2019, 31(4): 40-43. |
| 3 | DeGeorge KC, Holt HR, Hodges SC. Bladder cancer: diagnosis and treatment[J]. Am Fam Physician, 2017, 96(8): 507-514. |
| 4 | Burger M, Catto JW, Dalbagni G, et al. Epidemiology and risk factors of urothelial bladder cancer[J]. Eur Urol, 2013, 63(2): 234-241. |
| 5 | Godfrey DI, Le Nours J, Andrews DM, et al. Unconventional T cell targets for cancer immunotherapy[J]. Immunity, 2018, 48(3): 453-473. |
| 6 | Ward-Kavanagh LK, Lin WW, ?edy JR, et al. The TNF receptor superfamily in Co-stimulating and Co-inhibitory responses[J]. Immunity, 2016, 44(5): 1005-1019. |
| 7 | Janakiram M, Shah UA, Liu W, et al. The third group of the B7-CD28 immune checkpoint family: HHLA2, TMIGD2, B7x, and B7-H3[J]. Immunol Rev, 2017, 276(1): 26-39. |
| 8 | Wang Z, Wang Z, Zhang C, et al. Genetic and clinical characterization of B7-H3 (CD276) expression and epigenetic regulation in diffuse brain glioma[J]. Cancer Sci, 2018, 109(9): 2697-2705. |
| 9 | Kang FB, Wang L, Jia HC, et al. B7-H3 promotes aggression and invasion of hepatocellular carcinoma by targeting epithelial-to-mesenchymal transition via JAK2/STAT3/Slug signaling pathway[J]. Cancer Cell Int, 2015, 15:45. |
| 10 | 刘新波, 康富标, 曹娜娜, 等. B7-H3分子表达与食管癌复发转移的相关性研究[J]. 解放军医药杂志, 2019, 31(10): 23-26, 35. |
| 11 | Inamura K, Takazawa Y, Inoue Y, et al. Tumor B7-H3 (CD276) expression and survival in pancreatic cancer[J]. J Clin Med, 2018, 7(7):172. |
| 12 | Li YC, Guo GN, Song J, et al. B7-H3 promotes the migration and invasion of human bladder cancer cells via the PI3K/Akt/STAT3 signaling pathway[J]. J Cancer, 2017, 8(5): 816-824. |
| 13 | Xu ZL, Zhang Y, Wang L, et al. B7?H3 promotes malignant progression of muscle?invasive bladder cancer[J]. Oncol Rep, 2018, 40(5): 2722-2733. |
| 14 | Flem-Karlsen K, Fodstad ?, Tan M, et al. B7-H3 in cancer-beyond immune regulation[J]. Trends Cancer, 2018, 4(6): 401-404. |
| 15 | Zhong C, Tao B, Chen Y, et al. B7-H3 regulates glioma growth and cell invasion through a JAK2/STAT3/Slug-dependent signaling pathway[J]. Onco Targets Ther, 2020, 13:2215-2224. |
| 16 | Wang L, Zhang Q, Chen W, et al. B7-H3 is overexpressed in patients suffering osteosarcoma and associated with tumor aggressiveness and metastasis[J]. PloS one, 2013, 8(8):e70689. |
| 17 | Li DB, Wang J, Zhou J, et al. B7-H3 combats apoptosis induced by chemotherapy by delivering signals to pancreatic cancer cells[J]. Oncotarget, 2017, 8(43): 74856-74868. |
| 18 | Dawood S, Austin L, Cristofanilli M. Cancer stem cells: implications for cancer therapy[J]. Oncology (Williston Park), 2014, 28(12): 1101-1107, 1110. |
| 19 | Nassar D, Blanpain C. Cancer stem cells: basic concepts and therapeutic implications[J]. Annu Rev Pathol, 2016, 11: 47-76. |
| 20 | Vlashi E, Pajonk F. Cancer stem cells, cancer cell plasticity and radiation therapy[J]. Semin Cancer Biol, 2015, 31: 28-35. |
| 21 | Liu ZX, Zhang WL, Phillips JB, et al. Immunoregulatory protein B7-H3 regulates cancer stem cell enrichment and drug resistance through MVP-mediated MEK activation[J]. Oncogene, 2019, 38(1): 88-102. |
/
| 〈 |
|
〉 |